Molecular cloning and expression of hepatitis C virus core protein and production of monoclonal antibodies to the recombinant protein.
Asian Pac J Allergy Immunol
;
1996 Jun; 14(1): 31-41
Article
in English
| IMSEAR
| ID: sea-36759
ABSTRACT
The gene encoding nucleocapsid (core) protein of hepatitis C virus (HCV) was isolated from a Thai blood donor infected with HCV genotype 1b. The nucleotide sequence of this clone showed a high degree of homology to that of 4 HCV strains isolated from other Thai blood donors as well as that of the HCV prototypes of genotypes 1a, 1b, 2a and 3a. The entire region of the core gene was cloned into an expression plasmid pGEX-3X to be expressed as a fusion protein with glutathione-S-transferase (GST). E. coli transformants containing this plasmid did not express the fusion protein. However, GST-HCV core fusion protein could be produced when the core gene was truncated at the 3' end resulting in a gene encoding only the first 123 amino acid residues of the core protein. This fusion protein was insoluble in standard buffers, but could be solubilized by sarkosyl and thus subsequently purified using glutathione-Sepharose 4B. The purified fusion protein was immunogenic and could react with antibodies from blood donors infected with all genotypes of HCV found in Thailand. In addition, two murine hybridoma clones secreting monoclonal antibodies specific to the recombinant HCV core protein were produced. The purified HCV core protein and the monoclonal antibodies to the recombinant protein will be useful for developing assay systems for detecting anti-HCV antibodies and HCV antigen, respectively.
Full text:
Available
Index:
IMSEAR (South-East Asia)
Main subject:
Plasmids
/
Thailand
/
Recombinant Fusion Proteins
/
Recombinant Proteins
/
Humans
/
Molecular Sequence Data
/
Base Sequence
/
Viral Core Proteins
/
Polymerase Chain Reaction
/
Immunoenzyme Techniques
Country/Region as subject:
Asia
Language:
English
Journal:
Asian Pac J Allergy Immunol
Year:
1996
Type:
Article
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